Application of real-time PCR on the development of molecular markers and to evaluate critical aspects for olive oil authentication

Olive oil authentication using DNA-based markers is becoming very important. qRT-PCR was an efficient tool in investigating the utility of PCR primers for olive oil authentication allowing to discard primers with low PCR efficiency. It also allows investigating of the relative effectiveness among four DNA isolation methods and therefore qRT-PCR would be useful for further optimisation of the DNA extraction protocols. The number of target molecules for the amplification of 80 bp amplicons was higher than that of 200 bp. Therefore the amplicon size should be optimised for olive oil authentication since the higher the number of templates the greater the probability of successful amplification. On conclusion, qRT-PCR is a useful tool in the development of molecular markers for olive oil authentication and it should be used for the optimisation of critical parameters such as the amplicon size and the DNA extraction method.